Unit IMAGING TECHNOLOGIES
- Course
- Biotechnology
- Study-unit Code
- A000992
- Curriculum
- In all curricula
- Teacher
- Maurizio Mattarelli
- Teachers
-
- Maurizio Mattarelli
- Hours
- 42 ore - Maurizio Mattarelli
- CFU
- 6
- Course Regulation
- Coorte 2020
- Offered
- 2022/23
- Learning activities
- Affine/integrativa
- Area
- Attività formative affini o integrative
- Academic discipline
- FIS/03
- Type of study-unit
- Opzionale (Optional)
- Type of learning activities
- Attività formativa monodisciplinare
- Language of instruction
- Italian
- Contents
- Elements of electromagnetism, wave optics, and geometric optics for understanding imaging techniques.
Operating principle and application examples of the main microscopy techniques, relevant for biotechnological applications. - Reference texts
- Fundamentals of Light Microscopy and Electronic Imaging, 2nd Edition Douglas B. Murphy Michael W. Davidson (Wiley-Blackwell Ed). Also available in the electronic resources of the University (in particular on Proquest Ebook Central).
A university text of General Physics II (electromagnetism and optics).
For example: P. Mazzoldi, M. Nigro, C. Voices, Elements of Physics, Vol. 2, (Ed. EdiSES).
D. Halliday, R. Resnik, J. Walker, Fundamentals of Physics (Vol. 2), Ambrosiana Publishing House, Milan.
Lecture notes, articles and textbook chapters indicated therein (available in the University's online resources). - Educational objectives
- The main objective of the course is to provide students with the basic knowledge to understand the theoretical foundation and some experimental method of bio-imaging.
The main knowledge acquired will concern the principles and potential of some experimental methods of optical and spectroscopic imaging, electron and scanning probe microscopy .
The main skills that the course proposes to transmit are:
- know how to use the fundamental physical laws to understand the operating principle of modern imaging technologies, their potential and their limits.
- know how to use and analyze the results of the main imaging methods for relevant applications in the field of Biotechnology. - Prerequisites
- In order to understand the contents of the Study-unit, it is advisable to have acquired basic knowledge of Physics, such as those provided in the Study-unit Physics in the first year of the Biotechnology course.
- Teaching methods
- Lectures are provided on all the topics covered by the course and the analysis of experiments concerning optical microscopies, SEM, and spectroscopic imaging.
- Learning verification modality
- The assessment of the educational objectives of the course includes an oral exam lasting about 30 minutes. The exam will consist in the brief presentation of an imaging technique studied during the course (approximately 15 minutes). The presentation must include a description of the instrumentation, the physical mechanism of interaction, the ability of the technique in terms of resolution and an example of application in the biological field. In addition to possible requests for clarification or further information during the presentation, the exam will then continue with some more general questions.
The purpose of the test is to verify: 1) the acquired knowledge of the theoretical problems and of the various techniques and instruments studied during the course; 2) the acquisition of a critical capacity aimed at choosing the most suitable experimental technique for the study of the sample under examination; 3) the ability to communicate effectively and relevantly in oral form. - Extended program
- Electromagnetic fields and waves. Basics on the polarization of matter. Refractive index. Absorption.
Elements of geometric optics. Refraction and Reflection. Snell's Laws. Limit angle. Lenses and mirrors. Optical systems: eye, telescope, microscope. Optical aberration.
Coherent Waves. Young's experiment. Interference and diffraction. Airy disk and diffraction limit. Interference from thin films. Bragg's law.
Spectral analysis. Diffraction grating. Interferometer. Optical filters. Spectral resolution.
Photoelectric effect and quantum nature of radiation.
Light sources: Sun, incandescent lamps, gas lamps, LEDs. LASER.
Thermal detectors. Photomultipliers. Detector a semiconductor. Photodiodes. Spatial detectors (FPA, CCD, CMOS).
Hyperspectral reflectance imaging. Color: stimulus and perception. Diffuse and specular reflectance. Colorimetry. The human eye. Mapping strategies. Application to the study of vegetation. Devices. Spectral indices.
Optical microscopy. Elements of a combined microscope. Illumination. Lateral and axial spatial resolution. Abbe's limit. Rayleigh criterion. Contrast formation. Microscopy: bright field, dark field, phase contrast, polarized light. Fluorescence microscopy. Physical mechanism of fluorescence. Fluorescent molecules. Markers. Techniques: FRET, FRAP, FLIM. Confocal fluorescence microscopy.
Strategies for increasing the resolution: engineering of the scattering volume and non-linear excitation. Techniques: 4Pi, SIM, SMLM STED, TIRF, 2P, SNOM.
Vibrational Spectroscopy. Normal modes of vibration in molecules and solids. Infrared absorption. FTIR microscopy. Raman effect. Raman microscopy. Measuring equipment. Basics of Brillouin spectroscopy. Combined optical systems.
Electron microscopy. Wavelength of electrons. Instrumentation: sources, lenses, detectors. Electron-matter interaction. Secondary and backscattered electrons. EDX. Electronic spectroscopy. Spatial resolution. Systems: SEM, TEM, STEM. Preparation of biological samples for electron microscopy.
Scanning probe microscopy. Scanning tunneling microscope. Atomic force microscopy. Imaging Modes.
Digital images. Sampling. Nyquist criterion. Characteristics of detectors. MTF as a measure of the resolution of an imaging system. Image processing. Calibration. Digital filters. Segmentation. Dimensional reduction of multispectral images. Principal component analysis